RNA interference-mediated knockdown of a cytochrome P450, CYP6BG1, from the diamondback moth, Plutella xylostella, reduces larval resistance to permethrin.
We have previously reported that a cytochrome P450, CYP6BG1, from Plutella xylostella was found to be overexpressed in 4th instars of a permethrin resistant strain and inducible in the susceptible counterpart. The findings suggested potential involvement of CYP6BG1 in permethrin resistance, hence warranted a functional analysis. To assess the functional link of the gene to permethrin resistance, we adopted RNA interference-mediated gene silencing (RNAi) by dsRNA droplet feeding. Here, real time PCR analyses show that oral delivery of dsRNA can efficiently reduce the expression of CYP6BG1. Knockdown of CYP6BG1 transcript was evident in midgut and larval tissues enclosed in carcass. As a consequence of knockdown, a significant reduction in resistance of larvae fed CYP6BG1 dsRNA was observed after 24 and 48h of exposure to permethrin. In addition, CYP6BG1 dsRNA feeding to larvae led to reduced total P450 activities of microsomal preparations toward model substrates p-nitroanisole and benzyloxyresorufin. These results indicate that the overexpressed CYP6BG1 participate in enhanced metabolism of permethrin, thereby, resistance. The knockdown of a non-overexpressed P450, CYP6BF1v4, from the same resistant P. xylostella strain did not lead to changes in the level of resistance to permethrin, supporting further the specific involvement of CYP6BG1 in the resistance.
Bautista
Ma Anita M
MA
Laboratory of Applied Entomology, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Japan. maanita_bautista@yahoo.com
Miyata
Tadashi
T
Miura
Ken
K
Tanaka
Toshiharu
T
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004351:Drug Resistance; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D009036:Moths / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D026023:Permethrin / Q000494:pharmacology*; D034622:RNA Interference
D019476:Insect Proteins; D026023:Permethrin; D003577:Cytochrome P-450 Enzyme System